Antizyme Inhibitor

Crystal structure of the Antizyme inhibitor

 Each monomer consists of two domains: a TIM-like α/β-barrel domain (residues 45–280) and a modified Greek key  β-sheet domain (residues 8–44 and 281–435). Helices are colored in red and β sheets  in yellow.

A sequence alignment and structural comparison of mouse AzI crystallographic dimer to mouse, human, and trypanosome ODC (mODC, hODC, and tODC, respectively) show high sequence identity (~50%) and structural similarity between AzI and ODC monomers (RMSD values of 1.85 Å, 1.6 Å, and 1.5 Å, respectively). The structural comparison of mouse AzI crystallographic dimer (mAzI, cyan and blueviolet ) to mODC (PDB code 7odc, (red and lime ) is shown. Superposition of the interface of mAzI and mODC showing the inter-subunit variable loops (AzI residues 355–362 and 387–401). AzI loops are in black, and <font color='black'>ODC loops are in <font color='black'>yellow . The two AzI monomers (<font color='cyan'>cyan , <font color='blueviolet'>blueviolet ) have only <scene name='Antizyme_Inhibitor/Azi_odc1/1'>43 contacting residues (< 3.5 Å apart), while there are more contacts between the two monomers of hODC, <scene name='Antizyme_Inhibitor/Azi_odc1/2'>mODC (<font color='red'>red , <font color='lime'>lime ), and tODC (74, 83 and 69, respectively). Moreover, the surface area buried by the two mODC monomers is significantly larger than the one buried by the AzI monomers. These features explain a very weak crystallographic AzI dimer. The zipper, formed by conserved hydrophobic residues in mODC, stabilizes its dimeric structure. These residues involve F397(B), Y323(B), Y331(A), Y331(B), Y323(A), and F397(A) (the names of the chains are in brackets). The residue Y331 in the <scene name='Antizyme_Inhibitor/Azi_odc1/3'>ODC zipper is substituted by S329 in AzI and interferes with the formation of a similar zipper in AzI. Hence, in <scene name='Antizyme_Inhibitor/Azi_odc1/4'>mAzI this hydrophobic zipper is absent. Many residues, participating in the ODC interdimer interface interactions, are conserved among the ODCs from variuos organisms, but in AzI these residues are not conserved. Furthermore, the AzI conserved residues do not participate in interdimer interactions. For example, mODC possesses <scene name='Antizyme_Inhibitor/Azi_odc1/5'>two salt bridges (K169–D364 and D134–K294) stabilizing the ODC homodimer. In AzI, these 4 corresponding residues (<scene name='Antizyme_Inhibitor/Azi_odc1/6'>K169-D362 and D134-K291, respectively) are also present, but are too far apart to form a salt bridge. The two AzI monomers are positioned farther apart, in comparison ot ODC monomers, preventing the formation of interdimer interactions.

<scene name='Antizyme_Inhibitor/Binding_site/6'>Overlap of the AzI and mODC structures suggests that AzI does not bind PLP. <font color='black'>PLP is in <font color='black'>yellow, <font color='lime'>ODC residues D88, R154, R277, and Y389 are in lime , and the corresponding <font color='magenta'>AzI residues A88, H154, S274, and D387 are in magenta. Many of the residues participating in PLP-ODC binding are not conserved in AzI. These include D88A, R154H, R277S, D332E, and Y389D (ODC residue numbers follow the sequence of AzI). Notably, the absence of even one of these interactions, (e.g. ODC R277A mutant) results in a 100-fold decrease in PLP binding, a 50% drop in Kcat, and a 7-fold decrease in KM. </StructureSection>

3D structures of Antizyme Inhibitor
3btn – AzI – mouse

1zo0 – AzI – rat - NMR

Additional Resources
For additional information, see: Cancer

Reference
Shira Albeck, Orly Dym, Tamar Unger, Zohar Snapir, Zippy Bercovich and Chaim Kahana. Crystallographic and biochemical studies revealing the structural basis for antizyme inhibitor function. [http://www.ncbi.nlm.nih.gov/pubmed/18369191?ordinalpos=2&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_RVDocSum Protein Sci. 2008 May; 17(5): 793-802. Epub 2008 Mar 27.]